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Objective To investigate the effect and mechanism of transplanting Leydig-like cells from BMSCs to repair the testicular function of diethylhexyl phthalate (DEHP) infected mice. Methods Six C57 mice were randomly divided into two groups (3 each): control group (corn oil) and DEHP group [900 mg/(kg.d) DEHP mixed with corn oil]. Venous blood was taken from the eyeball 28 days after treatment to determine the serum testosterone level. One testis was stored in paraformaldehyde for HE and TUNEL staining, the other side of testis, after homogenization, was used to determine the changes of apoptotic proteins of caspase-3, caspase-8, caspase-9, Bcl-2 and Bax with Western blotting, the contents of MDA and GSH and the activities of SOD and GSH-Px were determined. Twenty-seven C57 mice were poisoned with DEHP for 28 days, and then randomly divided into Leydig-like cell transplant group, BMSCs transplant group and control group (9 each). BMSCs and Leydig-like cells from BMSCs were transplanted into the testis of mice in corresponding groups. Venous blood was taken from 3 mice of each group at 7th, 14th and 21st day after transplantation for determining the serum testosterone, and bilateral testis were obtained simultaneously to determine the contents of MDA and GSH and the activities of SOD and GSH-Px. The testis of mice was got 7th day after transplantation for HE and TUNEL staining, and the changes of apoptotic proteins of caspase-3, caspase-8, caspase-9, Bcl-2 and Bax were detected simultaneously. Results Compared with control group, the serum testosterone level decreased significantly, testicular spermatogenic cell disorder, tissue cells showed apoptosis, the serum levels of apoptotic protein caspase-3, caspase-8, caspase-9 and Bax increased obviously, while of Bcl-2 decreased markedly in testis of DEHP group, the content of MDA increased, and of GSH decreased, the activities of GSH-Px and SOD were inhibited with statistically significant difference (P<0.05 or P<0.01). Compared with control group, the serum testosterone level in Leydig-like cell transplantation group increased significantly at 7th, 14th and 21st day after testicular cell transplantation. The serum level of testosterone in BMSC transplantation group was higher than in control group at 14th and 21st day after transplantation. The content of MDA in testicular tissue decreased and of GSH increased,and the activities of GSH-Px and SOD recovered at 7th, 14th and 21st day after testicular cell transplantation (P<0.05 or P<0.01).The 7th day after transplantation, the apoptosis of testicular tissue was alleviated obviously in Leydig-like cell transplantation group and BMSC transplantation group than that in control group, the morphology of spermatogenic cells recovered, and the expression levels of apoptotic protein caspase-3, caspase-8, caspase-9 and Bax decreased significantly, while of Bcl-2 protein increased obviously(P<0.05 or P<0.01). Conclusion Transplantation of Leydig-like cells derived from BMSCs may repair the DEHP induced testicular damage by alleviating testicular oxidative stress and inhibiting testicular apoptosis, increase the teatosterone secretion, so as to promote the recovery of damaged testicular function.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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